Pharmacogenetics of Carboxylesterase 1

Carboxylesterases (CESs) catalyze the hydrolysis of a variety of ester- and amide containing compounds to their respective free acids. The main CES isozymes involved in drug metabolism are carboxylesterases 1 and 2 (CES1 and CES2). CES1 contributes to an estimated 80 to 95% of the total hydrolytic activity in the human liver. A CES1 c.428G>A (p.G143E, rs71647871) single nucleotide variation (SNV) markedly decreases the catalytic efficiency of CES1 in vitro. Possible effects of this variant on drug pharmacokinetics in vivo in humans, however, have not been systematically investigated. Furthermore, only a few studies have investigated the effects of CES1 variants on its expression and/or activity. Therefore, this thesis aimed to investigate genetic variability in the CES1 gene in the Finnish population and their effects on drug pharmacokinetics and pharmacodynamics in humans. The frequency distribution of the CES1 c.428G>A SNV was investigated in 860 healthy Finnish volunteers. The effects of the CES1 c.428G>A SNV on the pharmacokinetics of oseltamivir, clopidogrel, quinapril, and enalapril were investigated in 40 healthy volunteers. The CES1 gene and its flanking regions were sequenced in 192 healthy volunteers to identify previously unknown variants affecting CES1 whole blood expression. The findings were replicated in another set of 88 healthy volunteers. Furthermore, the effects of the detected variants on CES1 liver expression were investigated in 177 liver samples and on clopidogrel pharmacokinetics in 106 healthy volunteers from previous pharmacokinetic studies on clopidogrel. The CES1 c.428G>A variant allele was found with a minor allele frequency of 2.2%. The c.428G>A SNV reduced the hydrolysis of oseltamivir to the active oseltamivir carboxylate. The oseltamivir carboxylate to oseltamivir area under the plasma concentration-time curve (AUC) ratio was 23% smaller in heterozygous carriers than noncarriers. The c.428G>A SNV reduced the hydrolysis of clopidogrel to the inactive carboxylic acid metabolite. Consequently, the AUC of the parent clopidogrel was about 120% higher and that of the active metabolite about 70% higher in carriers than in noncarriers. Consistently, the c.428G>A SNV markedly enhanced the platelet inhibitory effect of clopidogrel. The average percentage inhibition of platelet aggregation at 0-12 hours was 19% higher in carriers than in noncarriers. The c.428G>A SNV significantly reduced the hydrolysis of enalapril to active enalaprilat. The AUC of enalaprilat was 20% lower in carriers than in noncarriers. The c.428G>A SNV had no observable effect on the pharmacokinetics of quinapril. Two intronic CES1 rs12443580 and rs8192935 SNVs were discovered to have a major effect on CES1 expression in whole blood, but not the liver. Moreover, these two SNVs had no effect on clopidogrel pharmacokinetics. In conclusion, the CES1 c.428G>A SNV reduces the bioactivation of oseltamivir, markedly increases the clopidogrel active metabolite plasma concentrations and antiplatelet effects, and reduces the bioactivation of enalapril in vivo in humans. The two intronic CES1 rs12443580 and rs8192935 SNVs have tissue-specific effects on CES1 expression. This could lead to substrate-dependent effects of these SNVs on drug biotransformation.Karboksyyliesteraasit hydrolysoivat erilaisia esteri- ja amidirakenteisia yhdisteitä. Lääkeaineiden hydrolyysiin osallistuvat lähinnä karboksyyliesteraasit 1 ja 2 (CES1 ja CES2). CES1 katalysoi noin 80-95 % maksassa tapahtuvasta hydrolyysistä. CES1-geenissä tunnetaan sen toimintaa in vitro heikentävä yhden nukleotidin muunnos (c.428G>A, p.G143E, rs71647871), mutta sen vaikutuksia lääkkeiden farmakokinetiikkaan ei ole aiemmin systemaattisesti tutkittu. Lisäksi muiden mahdollisten CES1-geenimuunnosten vaikutuksista CES1:n ilmentymiseen ja/tai aktiivisuuteen on vain vähän tietoa. Väitöskirjatyön tavoitteena oli tutkia CES1-geenimuunnosten esiintyvyyksiä suomalaisilla sekä näiden geenimuunnosten vaikutuksia CES1:n ilmentymiseen sekä lääkeaineiden farmakokinetiikkaan ja farmakodynamiikkaan ihmisillä. CES1 c.428G>A geenimuunnoksen esiintyvyyttä tutkittiin 860 terveen vapaaehtoisen koehenkilön otoksessa. CES1-geenimuunnoksen vaikutusta oseltamiviirin, klopidogreelin, kinapriilin ja enalapriilin farmakokinetiikkaan tutkittiin yhteensä 40 terveellä vapaaehtoisella koehenkilöllä. CES1-geeni sekvensoitiin 192 terveeltä vapaaehtoiselta koehenkilöltä mahdollisten CES1:n ilmentymiseen kokoveressä vaikuttavien geenimuunnosten löytämiseksi. Löydettyjen geenimuunnosten vaikutus toistettiin 88 terveen vapaaehtoisen aineistossa. Lisäksi geenimuunnosten vaikutusta tutkittiin CES1:n ilmentymiseen maksassa 177 maksanäytteen aineistossa ja klopidogreelin farmakokinetiikkaan 106 terveellä vapaaehtoisella koehenkilöllä aiemmin tehtyjä farmakokineettisiä töitä hyödyntäen. CES1 c.428G>A geenimuunnoksen esiintyvyys oli 2,2 %. CES1-geenimuunnos vähensi oseltamiviirin metaboliaa aktiiviseksi oseltamiviirikarboksylaatiksi, minkä seurauksena oseltamiviirikarboksylaatin ja oseltamiviirin pitoisuus-aikakäyrän alle jäävän pinta-alan (AUC) suhde oli 23 % pienempi kantajilla kuin ei-kantajilla. CES1-geenimuunnos vähensi klopidogreelin metaboliaa inaktiiviseksi karboksyylihappometaboliitiksi, minkä seurauksena klopidogreelin AUC oli noin 120 % suurempi ja aktiivisen metaboliitin AUC noin 70 % suurempi kantajilla kuin ei-kantajilla. Vastaavasti CES1-geenimuunnos lisäsi merkittävästi klopidogreelin antitromboottista vaikutusta; 12 tunnin keskimääräinen verihiutaleiden paakkuuntumisen esto oli 19 prosenttiyksikköä suurempi kantajilla kuin ei-kantajilla. CES1-geenimuunnos vähensi enalapriilin metaboliaa aktiiviseksi enalaprilaatiksi, minkä seurauksena enalaprilaatin AUC oli 20 % pienempi kantajilla kuin ei-kantajilla. CES1-geenimuunnoksella ei ollut vaikutusta kinapriilin farmakokinetiikkaan. Kaksi intronissa sijaitsevaa CES1-geenimuunnosta (rs12443580 ja rs8192935) vaikuttivat merkittävästi CES1:n ilmentymiseen kokoveressä, mutta eivät maksassa. Näillä kahdella geenimuunnoksella ei myöskään ollut vaikutusta klopidogreelin farmakokinetiikkaan. Yhteenvetona voidaan todeta, että CES1 c.428G>A geenimuunnos heikentää oseltamiviirin bioaktivaatiota, lisää klopidogreelin aktiivisen metaboliitin plasmapitoisuuksia ja antitromboottista vaikutusta sekä heikentää enalapriilin bioaktivaatiota. Kahden CES1-geenimuunnoksen (rs12443580 ja rs8192935) vaikutus CES1:n ilmentymiseen vaikuttaa olevan kudosspesifistä ja siten niiden vaikutus lääkeaineiden metaboliaan voi vaihdella eri aineiden välillä

How eSports players create brand loyalty through online engagement

Who would have known back in the late ‘50s, when the first video game was invented and video gaming phenomenon started, that in the 21st century it would be a major phenomenon, where hundreds of millions of players from all around the world are trying to win gaming tournaments and to get fame for itself. This phenomenon is called eSports. The main reason for this thesis is to get an idea of how eSports players create and could create brand loyalty for themselves through social media. This subject has never been studied before and since the gaming industry is growing every year, it is worth analyzing this phenomenon. The behavior of the fans in the online community is divided into different segments. The behavior of the streamers seems like having a major role in getting either positive or negative feedback from the fans. By owning more positive effects the players more likely to get more followers and fans, which in turn increases the player’s brand

Maarevisjonid Eesti- ja Liivimaal Rootsi võimuperioodi alguses

Land revisions in Estland and Livland at the beginning of the Swedish reign The article focuses on the ways land revisions were carried out in the provinces of Estland and Livland at the beginning of the Swedish reign and the purposes they served. Land revision data from the Swedish period has been used in numerous studies on Estonian and Latvian settlement and agrarian history. The current picture we have of the structure and changes of the population and settlements, enfeoff ments, imposts on peasantry, and agricultural conditions in various regions originates mainly from these sources. The materials on the revisions are kept in the Chamber Archives of the National Archives of Sweden, Historical Archives of the National Archives of Estonia, and the State Archives of Latvia. The revisions done within the territory of Estonia have been published which has notably expanded the scope of their use. The article pays special attention to the process of carrying out the revisions, temporal framework, methods, people who performed the tasks, but also to the general goals of Swedish economic rule in the Baltic provinces. There were several setbacks in conquering the area for the Swedish crown. Every time a part of the land became a Swedish possession, a swift assessment of the landed property was carried out. The Swedish crown organized several big revisions at the beginning of its rule in these provinces, namely in Estland from 1586–90 and in Estland and Livland from 1600–01, and additionally in Livland from 1624–27 and in 1638. Apart from these major revisions, there were several other limited revisions that were not completed or the results of which have only partially been preserved (from 1617–19 and in 1630). Revisions took place on the king’s command and according to specific instructions. Land revisions were primarily organized to plan, examine, and specify the state’s revenues. The tasks of the revision commission were generally rather broad, and one of the most important obligations during the first revisions was to check the landowners’ right of possession. It is especially evident during the revision in Estland from 1586–90. It was typical for Swedish revisions in Estland and Livland to be were carried out by commissions created for that purpose. On rare occasions, the revision was managed by the landlord visiting either Estland or Livland, as was the case with Duke Karl in 1601. The custom of delegating the organization of the revision to numerous sub-commissions was practiced in Livland in 1638. Members of the commission were typically appointed from the ranks of the high nobility of Sweden, Estland, and Livland. Mixed commissions of Swedes and Germans were common, but the local German members were naturally special trustees to the Swedish government. In 1638, the commissions consisted again mostly of local German officials, primarily lawyers. The real executer of the work was, however, a lower-level official (a secretary or scribe) appointed by the commission. It is clear that the knowledge of those officials about local conditions varied greatly, but the use of previous economic documentation and helpers, and the questioning of taskmasters and peasants improved the level of outcome. Intervals between revisions in Estland and Livland were very long, and every revision collected different information in comparison with the previous one. The state revenues from agriculture in the Baltics were often limited due to enfeoff ment, because only during the time of political turmoil and war and in their immediate aftermath were most of the farmsteads taxed by the state. The state representatives did not keep an account on the tax revenues of the enfeoffed territories. It sufficed when the officials checked from time to time whether the landed property did indeed belong to the nobles who claimed the relevant right. The revisions in Estland and Livland can be viewed in parallel. The major revisions in Estland took place from 1586–90 and again in 1601, and in Livland from 1624–27 and in 1638. The interval between the revisions was 11–12 years, so that the first revision was carried out during a longer period, while the later one only in the course of a single year. The first revisions were organized in the shadow of war, and the latter ones already during the time when Swedish reign was solidifi ed. The earlier revisions surveyed the territory that had only just been conquered by Sweden and the opportunities for enfeoff ment were still great. The latter revisions reflect the situation when the enfeoff ment had been almost accomplished. It is remarkable that aft er 1638 there were no revisions in Estland or Livland before the big cadastre of 1688–1690. The reason was that nearly all landed property had been enfeoff ed and the system of manors had become stable. KEYWORDS: Estonian history, province of Estland, province of Livland, history of Sweden, early modern period, agrarian history, payments, land revisions, manors, peasants. ÜLLE TARKIAINEN (b. 1957) is a Senior Researcher at the Institute of History and Archaeology, University of Tartu.130 Correspondence: Institute of History and Archaeology, University of Tartu, Ülikooli18, 50090 Tartu, Estonia. E-mail: [email protected]

Motivated, resistant, unfortunate and blameworthy – Positions constructed for Finnish long-term unemployed individuals through the employability discourse

In this study I examined the positions Finnish frontline workers constructed for their long-term unemployed clients through the employability discourse. The data consisted of 34 telephone interviews with rehabilitation counsellors working as part of a multi-sectoral team aiming to enhance the employability of unemployed jobseekers who face multiple barriers to attaining a job. In my analysis I identified four main positions: motivated, resistant, unfortunate and blameworthy. Within all of these positions, the responsibility related to employability enhancement was scrutinized. My analysis revealed that frontline workers position themselves as responsible or not responsible for enhancing their long-term unemployed clients’ employability depending on how employability or lack of it is constructed and a long-term unemployed individual positioned.Peer reviewe

Revisions to Client and Professional Self-Categorisations during Reciprocal Support Groups among the Long-Term Unemployed in Finland

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